Alternative dietary protein sources for Atlantic salmon (Salmo salar L.) effect on intestinal microbiota,intestinal and liver histology and growth

The present study has addressed the issue of dietary alterations on gut microbiota in fish by investigating modulation of the allochthonous and autochthonous bacterial gut community of Atlantic salmon following feeding with pea protein concentrate, soy protein concentrate, extracted sunflower, poultry by‐product and feather meal. The results revealed that total allochthonous and total autochthonous bacteria isolated from the distal intestine (DI) were affected by dietary modulations. Total autochthonous bacteria in proximal intestine remained unaffected. Corynebacteriaceae dominated the allochthonous bacteria and Lactobacillaceae dominated the autochthonous bacteria community in proximal and DI indicating that the most abundant groups are not shifted by alternative proteins. Some variations were observed between the bacterial communities as result of dietary manipulations. Alternative protein sources resulted in no severe signs of enteritis. Vacuolization of the liver was unaffected by diet. Furthermore, addition of plant materials significantly affected production of acetic acid. Weight of pyloric‐, proximal‐ and DI was influenced by diet, especially feather meal. Growth performance was significantly affected by inclusion of feather meal to the diet. In conclusion the results showed that pea protein concentrate and poultry by‐products are suitable protein replacements for Atlantic salmon and that soy protein concentrate is a potential candidate.     

The effect of dietary inulin on aerobic bacteria associated with hindgut of Arctic charr (Salvelinus alpinus L.)

The primary aim of the present study was to evaluate the population level of adherent (autochthonous) aerobic and facultative anaerobic bacteria in the hindgut of healthy Arctic charr (Salvelinus alpinus L.) fed dextrin or inulin. This was assessed by the dilution plate technique, and visualized using both transmission and scanning electron microscopy. A population level of 4.8 × 10⁵ adherent bacteria per gram wet mass was found in the hindgut of fish fed a casein‐based diet supplemented with 15% dextrin. However, substituting dextrin with 15% inulin reduced the bacterial population level in the hindgut (3.56 × 10⁴). A total of 217 bacterial isolates were identified by key phenotypical and biochemical characteristics. In addition, 22 strains were also identified by partial sequencing of the 16S rRNA gene. The composition of bacteria colonizing the hindgut of Arctic charr fed dextrin was dominated by the genera Staphylococcus, Pseudomonas, Micrococcus, Psychrobacter glacincola and Streptococcus. However, bacteria colonizing the hindgut of fish fed inulin were dominated by Gram‐positive bacteria of the genera Staphylococcus, Streptococcus, Carnobacterium and Bacillus. While Carnobacterium divergens‐like strains were isolated from charr fed dextrin, Carnobacterium maltaromicus‐like strains were isolated from the hindgut of fish fed inulin. Electron microscopical analysis of hindgut regions confirmed traditional culture‐based microbial analysis as fewer bacterial cells were observed between microvilli and associated with the surfaces of enterocytes of fish fed inulin rather than dextrin.     

Diversity of autochthonous bacterial communities in the intestinal mucosa of grass carp (Ctenopharyngodon idellus) (Valenciennes) determined by culture‐dependent and culture‐independent techniques

Traditional culture‐based technique and 16S rDNA sequencing method were used to investigate the mucosa‐associated autochthonous microbiota of grass carp (Ctenopharyngodon idellus). Twenty‐one phylotypes were detected from culturable microbiota, with Aeromonas, Shewanella, Lactococcus, Serratia, Brevibacillus, Delftia, Pseudomonas, Pantoea, Enterobacter, Buttiauxella and Yersinia as their closest relatives. Genomic DNA was directly extracted from the gut mucosa of C. idellus originating from six different geographical regions, and used to generate 609 random bacterial clones from six clone libraries and 99 archaeal clones from one library, which were grouped into 67 bacterial and four archaeal phylotypes. Sequence analysis revealed that the intestinal mucosa harboured a diversified bacterial microbiota, where Proteobacteria, Firmicutes and Bacteroidetes were dominant, followed by Actinobacteria, Verrucomicrobia and Deinococcus‐Thermus. The autochthonous bacterial communities in the gut mucosa of fish from different aquatic environments were not similar (C_s < 0.80), but γ‐Proteobacteria was a common bacterial class. In comparison to bacterial communities, the archaeal community obtained from one library consisted of Crenarchaeota and Euryarchaeota. These results demonstrate that molecular methods facilitate culture‐independent studies, and that fish gut mucosa harbours a larger bacterial diversity than previously recognized. The grass carp intestinal habitat selects for specific bacterial taxa despite pronounced differences in host environments.     

The effect of dietary krill supplementation on epithelium-associated bacteria in the hindgut of Atlantic salmon (Salmo salar L.): a microbial and electron microscopical study

Atlantic salmon (Salmo salar L.) were fed fishmeal protein for 46 days, and 500 g kg⁻¹ of fishmeal protein substituted with meal from Northern krill (Meganyctiphanes norvegica). No differences were observed in weight gain, length gain, feed conversion or specific growth rate between the groups that could be attributed to dietary manipulation. The adherent microbiota in the hindgut of the two rearing groups were further investigated. By substituting fishmeal with krillmeal, the total viable counts of aerobic and facultative aerobic bacteria colonizing the hindgut of Atlantic salmon increased from 8.5 × 10⁴ to 2.2 × 10⁶. Furthermore, dietary krillmeal affected the adherent hindgut microbiota. The Gram‐positive bacteria Carnobacteria piscicola, Microbacterium oxydans, Microbacterium luteolum and Staphylococcus equorum spp. linens and the Gram‐negatives Psychrobacter spp. and Psychrobacter glacincola were not isolated from hindgut of fish fed the krill diet. On the other hand, Pseudomonas fulgida, Pseudomonas reactans and Stenotrophomonas maltophila were not isolated from the control group fed fishmeal. Acinetobacter lwoffi, which is not normally found in the fish gut, was isolated from both feeding groups. Transmission electron microscopy showed bacteria‐like profiles between the hindgut microvilli in both feeding groups indicating autochthonous microbiota. When fish were fed the krill diet, hindgut enterocytes were replete with numerous irregular vacuoles. These vacuoles were not observed in fish fed the fishmeal protein.     

Characterisation of the microbiota associated with intestine of Atlantic cod (Gadus morhua L.): The effect of fish meal, standard soybean meal and a bioprocessed soybean meal

Populations of aerobic heterotrophic bacteria present in the gastrointestinal (GI) tract of healthy Atlantic cod (Gadus morhua L.) fed three different diets, fish meal, standard or a bioprocessed soybean meal (BPSBM), were estimated using the dilution plate technique. A total of 944 isolates were characterised by biochemical and physiological properties and 425 isolates were identified further by sequencing the 16S rRNA genes. Our results showed that gut microbiota were affected by dietary manipulation. The GI tract of fish fed fish meal was dominated by Gram-positive bacteria of the genera Brochothrix and Carnobacterium. The Gram-negative bacteria Chryseobacterium spp. and Psychrobacter glacincola, and Gram-positive bacteria belonging to Carnobacterium, dominated in the digestive tract of fish fed soybean meal. In contrast to these results, genus Psychrobacter dominated in the GI tract when fish were fed BPSBM.Until recently, it was generally suggested that the gut microbiota of fish were less diverse than in homoeothermic animals. However, the present study identified several “new” bacterial species isolated from the alimentary tract of Atlantic cod. These “new” bacterial species are not normally isolated from the GI tract of fish. Based on our finding we suggest that the GI tract microbiota of fish might not be as simple as believed.Antagonistic activity of carnobacteria regarding inhibition of growth of two fish pathogens (Aeromonas salmonicida ssp. salmonicida and Vibrio anguillarum) was observed. However, some difference in the antibacterial activity of Carnobacterium spp. was observed. Whether this antagonistic activity has any effect in challenge studies will be discussed, especially in relation to the finding that the digestive tract is one of the major infection routes in fish.     

Effect of intraperitoneal injection of immunostimulatory substances on allochthonous gut microbiota of Atlantic salmon (Salmo salar L.) determined using denaturing gradient gel electrophoresis

The allochthonous microbiota in the proximal and distal intestine was investigated in three groups of Atlantic salmon (Salmo salar L.) fed a commercial diet and intraperitoneally injected with (a) phosphate‐buffered saline (control), (b) lipopolysaccharide (LPS) from the fish pathogenic bacteria, Aeromonas salmonicida ssp. salmonicida, and (c) laminaran [β‐(1,3)‐d ‐glucan]. Denaturing gradient gel electrophoresis (DGGE) of the hyper variable V3 region was used to present the microbiota in different experimental groups. Sequencing and phylogenetic analysis of excised DGGE bands suggested that an intraperitoneal injection of LPS from A. salmonicida affects the allochthonous gut bacteria of Atlantic salmon to some extent, as Aeromonas enteropelogenes, Aeromonas veroni, Psychrobacter, Lactobacillus letvazi, Lactobacillus satsumensis, Pantoea, swine manure bacterium and several uncultured bacteria were unique for this group. On the other hand, the bacterial diversity of the allochthonous microbiota did not seem to be affected by injection of β‐(1,3)‐d ‐glucan. Sequences of this experimental group were most closely related to A. enteropelogenes, uncultured Escherichia and Lactobacillus aviarius ssp. aviarius.     

Supplementing enzymes to extruded,soybean‐based diet improves breakdown of non‐starch polysaccharides in rainbow trout (Oncorhynchus mykiss)

Plant‐based feed ingredients typically contain remnants of dietary fibres [DF; non‐starch polysaccharides (NSP) and lignin] that have various antinutritive effects in carnivorous fish. Exogenous enzymes have been shown to improve the apparent digestibility coefficients (ADC) of plant‐based diets presumably by assisting in the breakdown of NSP. This study examined the effects on NSP degradation when supplementing β‐glucanase, xylanase, protease or a mix of the three enzymes to an extruded, juvenile rainbow trout (Oncorhynchus mykiss) diet containing 344 g kg^?1 de‐hulled, solvent‐extracted soybean meal (SBM). The NSP content in the non‐supplemented control diet and in faecal samples from the dietary treatment groups was analysed to determine the recovery/apparent digestibility of cellulose and total non‐cellulosic polysaccharide (T‐NCP) sugar monomers. The enzymes had significant, positive effects at the pH range and temperature prevailing in the gastrointestinal tract: β‐glucanase improved the ADC of mannose, galactose and uronic acids; xylanase and protease improved the ADC of xylose; and protease furthermore improved the ADC of mannose and uronic acids. There were no effects when supplementing all three enzymes together. In conclusion, exogenous enzymes may potentially be applied to fish feed containing SBM, assisting in the breakdown of NSP and alleviating some of the antinutritive effects.     

Analysis of bacterial community and bacterial nutritional enzyme activity associated with the digestive tract of wild Chilean octopus (Octopus mimus Gould, 1852)

Information on the bacterial community associated with octopus is very scarce, unlike fish and other molluscs. This study revealed the bacterial community associated with digestive tract of wild Chilean octopus Octopus mimus using a culture‐dependent method and 16S rDNA clone library. Moreover, we analysed the bacterial nutritional enzyme activity of culturable bacteria. A culture‐dependent method showed that the composition of the culturable bacterial community was substantially different between female and male octopus. The predominant species in female octopus were Vibrionaceae and Streptococcaceae, whereas only Vibrionaceae was dominated in male octopus. Bacterial nutritional enzyme activities of culturable bacteria from male octopus were much higher than female octopus. The 16S rDNA clone library analysis showed that the bacterial community of male octopus exhibited a higher diversity than that of female octopus. The genus Mycoplasma was the predominant bacteria in the digestive tract of all octopus samples. The results obtained in this study raise the possibility that each octopus has different food consumption due to different bacterial community and nutritional enzyme activity, although Mycoplasma sp. is one of the predominant bacteria in the digestive tract. Moreover, our results are useful for the future of microbiological investigation associated with the octopus and for probiotics in the octopus aquaculture.     

Antimicrobial activity in bacteria isolated from Senegalese sole, Solea senegalensis, fed with natural prey

Intestinal bacteria in marine fish may produce antimicrobial substances which inhibit pathogenic bacteria. The aim of this study was to determine the influence of a change of fish diet on the antimicrobial activity of the culturable aerobic gut microflora of Senegalese sole, Solea senegalensis. Pre‐adult 15‐month‐old fish previously fed on an artificial diet, were fed polychaetes (Hediste diversicolor), which form part of the natural diet of Senegalese sole. Samples were taken0, 3 and 6 weeks after start of the experiment from the stomach, small and large intestine of the fish. The bacterial strains isolated from these samples were sub‐cultured to pure cultures and stored at ?80°C. Several biochemical tests were run to obtain some basic phenotypic characteristics of the isolated strains. Amplification and sequencing of 16S rDNA fragments were used to identify the majority of the bacterial strains isolated. The identification by use of this molecular approach gave results in agreement to the phenotypic characterization. Feeding with polychaetes significantly increased (P<0.05) the numbers of presumptive Vibrio isolates in the gut. The number of bacterial strains with antimicrobial activity, as determined by two in vitro approaches, was significantly (P<0.05) increased by feeding with polychaetes.     

The effects of oligofructose on growth performance,survival and autochthonous intestinal microbiota of beluga (Huso huso) juveniles

Preliminary experiments were undertaken to investigate the effects of oligofructose on beluga sturgeon (Huso huso) growth performance, survival and culturable autochthonous intestinal microbiota. Juveniles (20 g) were fed diets containing varying levels of oligofructose (10, 20 and 30 g kg^?1) at 2–3% body weight per day for 7 weeks. Compared to the control group, no significant (P > 0.05) effect on growth performance was observed in fish fed diets supplemented with oligofructose at 10 and 20 g kg^?1. However, compared to the 20 g kg^?1 group, feeding oligofructose at 30 g kg^?1 resulted in adverse effects on growth performance. Dietary supplementation of oligofructose at 20 g kg^?1 significantly increased survival rate. Microbiological assessment indicated that the viable culturable autochthonous levels were not affected by dietary oligofructose. Although lactic acid bacteria (LAB) were not a dominant component of the endogenous autochthonous microbiota, LAB levels were significantly elevated in fish fed 20 g kg^?1 dietary oligofructose. This elevated LAB population was able to persist for at least 1 week after reverting the prebiotic group back to a control diet. This study encourages further research on different aspects of oligofructose in sturgeon culture with clear emphasis on optimizing dosage levels.     

A study on bacteria associated with the intestinal tract of farmed yellow seahorse,Hippocampus kuda (Bleeker, 1852): characterization and extracellular enzymes

The microenvironment of bacteria associated with the gastrointestinal tract of an animal influences the host in many ways, including the metabolism of several nutrients. Isolation, molecular characterization and enzymatic activities of culturable bacteria associated with the gastrointestinal tract of hatchery‐reared oceanic yellow seahorse, Hippocampus kuda, were investigated in the present study. 16S rRNA sequencing of bacterial isolates yielded 10 different bacterial genera belonging to two major groups: proteobacteria (62.9%) and firmicutes (37.03%). The predominant microbial flora belonged to the genera Vibrio spp., Enterovibrio sp. and Bacillus spp. Burkholderia cenocepacia, hitherto unreported from the gastrointestinal tract of marine fish, was retrieved from the seahorse gut, albeit in small numbers. Enzymatic studies indicated that the bacterial isolates possess the ability to degrade lipids, cellulose, xylan, starch and proteins. Lipase activity was exhibited by a majority of the isolates (70.4%), indicating that the bacterial flora associated with the intestinal tract of seahorses is capable of digesting foods rich in lipids. The possible implications for the role of these bacteria in the digestion and physiological processes are discussed. To our knowledge, this is the first time that the characterization and enzymatic activities of bacteria associated with the intestinal tract of farmed seahorses have been reported.     

Quantitative and Qualitative Aspects of Bacterial Communities Associated with Cultures of Chlorella minutissima

Microbiological conditions in 200 L cultures of microalgae Chlorella minutissima in polyethylene sleeves were examined. The influence of addition of antibiotic (nitrofurantoin) was studied. Samples were taken 2, 5, 9, and 14 d after inoculation and were spread on solid Zobell medium and on thiosulfate citrate bile salts sucrose agar for the enumeration of total numbers of bacteria and numbers of presumptive Vibrio bacteria, respectively. Microalgae grew well in the cultures and high numbers of culturable bacteria (about 10⁷ bacteria/mL) were present in all microalgae cultures. Addition of antibiotic in 200‐L cultures of C. minutissima did not result in a significant decrease of total counts of bacteria per unit volume (P > 0.05), while the numbers of presumptive Vibrio were higher in cultures added antibiotic compared with the control treatment (P < 0.05). The numbers of presumptive Vibrio per unit volume and the percentage of fast‐growing bacteria were higher in microalgae supernatant than in noncentrifuged samples. Representative colonies were taken from all samples and in total 649 bacterial strains were isolated during this study. A range of phylotypes was identified by amplification and sequencing of the 16S rDNA gene and phylogenetic trees were constructed. Several members of the Roseobacter clade were dominant among the culturable isolates.     

Antagonism against fish pathogens by cellular components and verification of probiotic properties in autochthonous bacteria isolated from the gut of an Indian major carp,Catla catla (Hamilton)

Probiotic potential of the autochthonous bacteria in catla, Catla catla has been evaluated through determination of antagonistic activity (in vitro) of the cellular components of gut bacteria against seven fish pathogens. Altogether 208 strains were isolated, inhibitory activity of the isolates was evaluated through cross‐streaking and 16 primarily selected antagonistic strains were confirmed using the double‐layer method. Four bacteria that showed antagonism against ≥4 pathogens were selected as putative probiotics. The intracellular, extracellular, whole‐cell and heat‐killed cell components exhibited bactericidal activity against the pathogens. In addition, the selected strains were capable of producing different extracellular enzymes, competent to grow in intestinal mucus and could tolerate diluted bile juice. Analysis of 16SrRNA partial gene sequence revealed that both the strains CC1FG2 and CC1FG4 were Bacillus methylotrophicus (KF559344 and KF559345), while the isolates CC1HG5 and CC2HG7 were Bacillus subtilis subsp. spizizenii (KF559346) and Enterobacter hormaechei (KF559347) respectively. Bio‐safety evaluation through intra‐peritoneal injection of the isolates did not induce any pathological signs or mortalities in C. catla. The study confirmed probiotic properties of autochthonous gut bacteria in C. catla and demonstrated potential for using them as bio‐control agents. However, in vivo studies are essential to explore their efficacy in the commercial aquaculture.     

Use of macroalgae supplemented with probiotics in the Haliotis rufescens (Swainson, 1822) culture in Northern Chile

In this study, we evaluate the use of macroalgae as vectors of probiotics bacteria into the digestive tract of abalone to improve their survival and growth. It is shown that when abalone Haliotis rufescens of different sizes were fed with a natural diet composed of fronds of the macroalga Macrocystis integrifolia supplemented with a mixture of Vibrio sp. C21‐UMA, Agarivorans albus F1‐UMA and Vibrio sp. F15‐UMA bacteria, there was a significant increase (P<0.05) in the average monthly growth rate and survival (%)in a period of 210 days, compared with the control without a probiotic supplement. The permanence of the probiotics in the digestive tract of the animals was monitored, and it was found that the number of culturable C21‐UMA and F1‐UMA bacteria decreased significantly (P<0.05) in recently weaned and adult abalone, and they almost disappeared completely on day 19 of the bioassay. However, the culturable Vibrio sp. F15‐UMA disappeared completely from the digestive tract on day 22 of the bioassay, and these were the bacteria that remained at the highest concentration compared with the other two bacterial strains over the experimental period. It is therefore shown that it is feasible to use a probiotic mixture to improve the profitability of the H. rufescens culture.     

The autochthonous microbiota of the freshwater omnivores jundiá (Rhamdia quelen) and tilapia (Oreochromis niloticus) and the effect of dietary carbohydrates

Previous studies have shown that the gut microbiota of fish differs depending on host feeding habits and these populations may also be influenced by dietary ingredients. In this study, the bacterial populations of the intestinal tract of the opportunistic omnivore jundiá Rhamdia quelen and the typical omnivore tilapia Oreochromis niloticus were investigated in two experiments. In experiment I, the levels of amylolytic, cellulolytic, lipolytic, proteolytic and total culturable bacteria were compared in the proximal, mid and distal intestine using selective agar. Higher levels of amylolytic, cellulolytic, lipolytic, proteolytic and total culturable bacteria were observed in the tilapia than jundiá, in all intestinal regions. Jundiá contained higher levels of proteolytic bacteria and lipid digesting bacteria in the distal intestinal portion as compared with the mid intestine; both fish species also presented more amylolytic bacteria in the distal intestine. In experiment II, the amylolytic intestinal microbiota between the two species was compared after administration of diets containing different carbohydrate sources. Jundiá fed broken rice presented higher total culturable bacterial levels; however, dietary cassava bagasse and ground corn significantly elevated the population of amylolytic bacteria in tilapia (P > 0.05). PCR‐DGGE was also used to assess the bacterial communities in experiment II. A Cetobacterium spp. was detected in jundiá fed diets containing broken rice, and tilapia fed cassava bagasse, dextrin, broken rice and ground corn. Microbial differentiation was further demonstrated between jundiá and tilapia, because an uncultured bacterium was unique in tilapia and an uncultured spirochete was observed only in jundiá; the presence of these bacterial species was also influenced by dietary carbohydrate sources.     

Identification and partial characterization of potential probiotic lactic acid bacteria in freshwater Labeo rohita and Cirrhinus mrigala

Carps are the most diversified freshwater fish belonging to family Cyprinidae. Numerous probiotic and pathogenic lactic acid bacteria (LAB) have been characterized from carps. However, the diversity of these ecologically important bacteria is entirely unknown in freshwater fish of Pakistan. The present study aimed to characterize and identify the lactic acid bacteria from two carps viz. Laboe rohita and Cirrhinus mrigala and determine their antagonistic activity. Seventeen bacterial isolates were purified from the gastrointestinal tract and gills of these fish and characterized morphologically. Initially, seven isolates were screened as LAB using agar supplemented with CaCO₃. Subsequently, only two isolates CILB2 and RIL10 were selected as LAB after high‐performance liquid chromatography analysis for lactic acid production. Isolates CILB2 and RIL10 were genetically identified as Enterococcus faecalis and Weissella sp., respectively after 16S rRNA gene sequence analysis. Both strains exhibited significant antagonistic activity against common fish pathogens Streptococcus agalactiae, Staphylococcus aureus and Escherichia coli. Enterococcus faecalis CILB2 and Weissella sp. RIL10 were also found negative for haemolysis and gelatinase activities and were sensitive to ampicillin, amoxicillin, doxycycline, erythromycin, chloramphenicol and co‐trimoxazole antibiotics. The identified LAB strains may further be investigated for their potential probiotic application in fish feed and food preservation techniques.     

The effect of dietary chitin on the autochthonous gut bacteria of Atlantic cod ( Gadus morhua L.)

In the present study the impact on autochthonous (adherent) bacteria in proximal intestine (PI) and distal intestine (DI) of Atlantic cod ( Gadus morhua L.) was evaluated following feeding of a control diet and a diet supplemented with 5% chitin. The autochthonous gut bacteria were investigated using denaturing gradient gel electrophoresis (DGGE). Analysis of the microbiota associated with PI and DI of Atlantic cod indicate that dietary chitin modulate the intestinal bacterial community. For example, band 25 ( Escherichia coli–like), band 14 ( Anaerorhabdus furcosa–like) and band 29 (uncultured bacterium–like) in PI were depressed by dietary chitin (P < 0.05). The number of bands (23.7 ± 5.4) in DI of fish fed chitin was marginally higher than the control fish (16.7 ± 2.1) (P = 0.065), and the relative abundances of band 6 (swine faecal bacterium–like) were marginally stimulated by dietary chitin (P = 0.095). Furthermore, the present study reports several novel sequences not previously reported in the gastrointestinal tract of Atlantic cod. Whether the dietary effect of chitin on gut bacterial community has any positive effect of fish health merits further investigations.     

The effects of soybean, linseed and marine oils on aerobic gut microbiota of Arctic charr Salvelinus alpinus L. before and after challenge with Aeromonas salmonicida ssp. salmonicida

Populations of heterotrophic bacteria present in the hindgut region of Arctic charr Salvelinus alpinus L. fed dietary soybean, linseed and marine oils before challenge with Aeromonas salmonicida ssp. salmonicida and marine oil after challenge were estimated using the dilution plate technique. There were differences in bacterial composition between the rearing groups before and after challenge, as well as interindividual variations. For example, carnobacteria were only isolated from the hindgut region of fish fed soybean oil and linseed oil before challenge, whereas Carnobacterium spp. and Carnobacterium funditum‐like species were isolated from fish fed the same oils after challenge. Three non‐motile Aeromonas spp. were isolated from infected fish fed marine oil. One of these isolates was identified as identical to A. salmonicida ssp. salmonicida used in&the challenge test by microbial fingerprinting (amplified fragment length polymorphism). Electron microscopic examinations of hindgut regions demonstrated substantial numbers of bacterial cells associated with enterocytes, but bacterial colonization of the enterocyte surface varied between different rearing groups. The potential of bacteria found associated with the hindgut region to inhibit the fish pathogens A. salmonicida, Vibrio salmonicida and Vibrio anguillarum differed between rearing groups.     

Effects of prebiotics on nutrient digestibility of a soybean‐meal‐based diet by red drum Sciaenops ocellatus (Linnaeus)

The present study examined the effects of four prebiotic compounds on nutrient and energy digestibility of soybean‐meal‐based diets by red drum (Sciaenops ocellatus). The experimental diets contained 40% crude protein of which approximately half was provided by soybean meal with the remainder from menhaden fish meal. The four prebiotics GroBiotic^®‐A (a mixture of partially autolysed brewers yeast, dairy ingredient components and dried fermentation products), mannanoligosaccharide (MOS), galacto‐oligosaccharide (GOS) and inulin were individually added to the basal diet at 1% by weight. A diet with all its protein provided by menhaden fish meal was also included as a control. This control diet had the highest apparent digestibility coefficient (ADC) values – 87% for protein, 87% for lipid, 78% for organic matter and 83% for energy. The basal soybean‐meal‐based diet supplemented with GroBiotic^®‐A, GOS and MOS had significantly (P<0.05) increased protein (82%, 82%, 82% respectively) and organic matter ADC values (69%, 64%, 66% respectively), compared with the basal diet (69% for protein and 49% for organic matter). However, the lipid ADC values were significantly decreased for fish fed with the diets supplemented with inulin, GOS and MOS (63%, 61%, 61% respectively) compared with the basal diet (77%) but not for those fed GroBiotic^®‐A (82%). Energy ADC values were also increased in fish fed with the GroBiotic‐A^®, GOS and MOS diets (73%, 70%, 72%), compared with the basal diet (57%); however, fish fed with the inulin diet had an energy ADC value (54%) similar to that of fish fed with the basal diet. Thus, the present study is the first to demonstrate that nutrient and energy digestibility of soybean‐meal‐based diets by red drum can be enhanced with prebiotic supplementation.     

Growth and intestinal morphology in cobia (Rachycentron canadum) fed extruded diets with two types of soybean meal partly replacing fish meal

Juvenile cobias, Rachycentron canadum, were fed extruded diets containing toasted defatted soybean meal (SBM) or untoasted defatted SBM [white flakes (WF)] to study growth and feed conversion, and to study if SBM induces morphological changes in the gastrointestinal (GI) tract. Three diets were produced: a fish meal‐based control diet (FM diet) with 558 g FM kg^?1, and two diets with 335 g FM and either 285 g SBM kg^?1 (SBM diet) or 285 g WF kg^?1 (WF diet). The diets were extruded at approximately 120°C with 280 g kg^?1 moisture. Triplicate groups of cobias (mean weight: 25.9 g) were fed the diets during 6 weeks. Feed intake of the FM and SBM diets were not significantly different, whereas the cumulative feed intake of cobias fed the WF diet was lower (P < 0.05) than that of cobias fed the FM and SBM diets after the first 21‐day period. Specific growth rate and feed conversion ratio were not significantly different between cobias fed the FM and SBM diets, but significantly poorer results were obtained in cobias fed the WF diet. No morphological differences in the GI tract could be attributed to the diets, and cobias fed soy did not develop enteritis in the distal intestine.